Study On Immobilization Of Recombinant Chitinase With Two Functional Magnetic Carriers

Magnetic polymer microsphere is a new type of functional polymer material that has been developed recently. Because of its good magnetic responsiveness, small size, large specific surface area and good biocompatibility, it is widely used in various areas of biotechnology. Two kinds of functionalized magnetic carriers(A-type, P-type) were prepared and assessed their application in immobilizing recombinant chitinase in this paper, and researched immobilized rechitinase.Fe3O4magnetic particle was synthesized by the modified chemical co-precipitation method, ferrous chloride, ferric chloride, sodium hydroxide as the main material. The average diamete of Fe3O4magnetic particle were well, the size distribution and dispersion uniformity were well. There had the characteristic absorption peaks in the FTIR, corresponding to the Fe-O and-OH.Two kinds of functionalized magnetic carriers (A-type, P-type) were prepared in this paper.The A-type carrier was prepared by3-aminopropyltriethoxysilane (APTES) and Fe3O4magnetic microparticle, the P-type carrier was prepared by copolymerization of Glycidyl methacrylate (GMA) on the surface of Fe3O4magnetic microparticle.The A-type magnetic microsphere diameter was about100nm. It was superparamagnetic and-NH2groups coated on the surfae.The P-type microsphere diameter was about100nm, approached the spherical. There had the characteristic absorption peaks of GMA epoxy molecules in the FTIR.A-type and P-type were used to immobilize recombinant chitinase and assessed their optimal conditions of immobilizing recombinant chitinase in this paper. The results show that, the optimum condition for A-type were:the amount of enzyme to take100mL, the glutaraldehyde concentration11%50mL, the reaetion time5h. The optimum condition for P-type were:the amount of enzyme to take120mL, the reaetion time9h.Studied on the two kinds of functional magnetic carriers fixed effect, and the nature of the immobilized enzyme.The results show that P-type carrier has higher protein loading capacity compared with A-type carrier,but the two carriers had the similiar purification multiple.It was prove that the functional magnetic carriers could enhanced the storage stability,the thermo stability, the pH endurance compared to the crude extract.The immobilized recombinant chitinase were more than44%of the original activity even reusing up to seven times. The optimum pH range of immobilized recombinant chitinase was6.8-8.